An Immunological Assay for the Sigma Subunit of RNA Polymerase in Extracts of Vegetative and Sporulating Bacillus subtilis

AUTOR(ES)

Tjian, Robert

RESUMO

The activity of the σ subunit of Bacillus subtilis RNA polymerase decreases markedly during the first hours of sporulation [T.G. Linn et al. (1973) Proc. Nat. Acad. Sci. USA 70, 1865-1869]. We have prepared antibody against RNA polymerase holoenzyme to determine the fate of σ polypeptide during spore formation. This antiserum specifically and independently precipitates σ and core polymerase from crude extracts of B. subtilis as judged by both sodium dodecyl sulfate and urea gel electrophoresis of the precipitates. We report that crude extracts of sporulating cells lacking σ activity contain as much σ polypeptide as extracts of vegetative cells. However, σ polypeptide in extracts from sporulating cells is apparently only weakly associated with RNA polymerase, as indicated by the failure of σ to co-purify efficiently with core enzyme during phase partitioning.

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